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BioMed Research International
Volume 2014, Article ID 851349, 19 pages
Research Article

Atg6/UVRAG/Vps34-Containing Lipid Kinase Complex Is Required for Receptor Downregulation through Endolysosomal Degradation and Epithelial Polarity during Drosophila Wing Development

Department of Anatomy, Cell and Developmental Biology, Eotvos Lorand University, Budapest 1117, Hungary

Received 24 January 2014; Accepted 1 April 2014; Published 21 May 2014

Academic Editor: Gábor Juhász

Copyright © 2014 Péter Lőrincz et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Fly stocks used in this study are listed in Table S1.

Antibodies used in this study are listed in Table S2.

The details and exact P values of statistical analyses are listed in Table S3.

The genotypes of the examined animals are listed in Table S4.

Figure S1: Impaired endocytic degradation of NICD and Delta in Atg6 depleted cells. Imaginal wing discs expressing Atg6 dsRNA by enGal4 were immunostained against (a) the intracellular domain of Notch (NICD) and (c) Delta. The region of RNAi is marked by the coexpression of GFP and the borderline of the region is indicated with a dashed white line. (a,c) In Atg6 RNAi cells both NICD and Delta accumulated in small intracellular dots. (d,b) Quantification of a,c. On box plots, bars show the data ranging between the upper and lower quartiles; median of the signal covered areas is indicated as a horizontal black line within the box. Whiskers plot the smallest and largest observations, while dots and asterisks indicate outliers. NS means P>0.05, *** means P<0.001. For details and exact p values of statistical analyses see Table S3. For genotype see Table S4. Scale bars represent 50 μm.

Figure S2: The penetrance of the wing phenotype of RNAi lines. Cross indicates increased lethality. See Figure 9 for phenotypes.

Figure S3: Cell death in Atg14 RNAi wing discs. (a-d) Imaginal wing discs expressing RNAi in the wing pouch by BxMS1094Gal4 were immunostained against Wnt. In contrast to (a) controls, in the (b) Atg14 RNAi wing disc numerous cleaved Caspase-3 positive picnotic cells could be observed in the wing pouch (arrows). RNAi of (c) Atg6 or (d) UVRAG did not result in the appearance of cleaved Caspase-3 positive cells in the wing disc. (e-h) TUNEL assay in wing discs. Compared to (e) control or (g) Atg6 RNAi or (h) UVRAG RNAi discs, the wing pouch of (f) Atg14 RNAi discs contained numerous TUNEL-positive cells (arrows), indicating apoptosis. For genotypes see Table S4. Scale bars represent 50 μm.

  1. Supplementary Tables
  2. Supplementary Figures